The ultimate goal of the project is to de novo design and synthesize transmembrane proton transporters for biomedical applications. My approach is to learn from nature and to systematically study and reproduce components of the more complex natural proton pumps. The first part of the work will be focused on mechanism of the vectorial proton movement. Common characteristic of several natural membrane pumps is the essential conformational change that leads to the displacement of proton across a dielectric barrier. The energy input that causes the conformation change must be a tight choreography of several functions: pK shifts, barrier crossing motion and proton binding and release. Under the NRSA fellowship I will reproduce these functions within one, more simple, model protein (maquette). As progress will be made, I will contribute rich supply of information essential to understanding of principles underlying these fundamental natural protein functions and develop essential guidelines for engineering for function. The second step toward our goal is engineering of proton channel maquettes. Two strategies will be used: searching for and using obvious functional consensus motifs within natural proton pumps and utilizing considerable experience drawn from de novo design of already fabricated proton channels. The proton pump maquettes will be engineered for incorporation into the hydrophobic core of natural as well as synthetic membranes to obtain robust systems that are required for many biotechnology/clinical applications.